title: Optimization of recombinant hexaoligochitin-producing chitinase production with response surface methodology.
abstract: Hexaoligochitin produced by chitinase, ASCHI61, from Aeromonas schubertii was recently expressed. In this work, the optimal conditions for the mass production of ASCHI61 were investigated. The efficiency of recombinant protein expression in Escherichia coli was determined by various parameters, including the pH of the culture medium, induction temperature, shaking speed, inducer concentration, and induction period. The optimization experiments could be simplified through a statistical design of experiments (response surface methodology). From the fractional factorial design, the interactive effect of induction temperature and time was the most significant. The total activity of the enzyme was 32,092 U at 23.9 °C with 115 min of induction. Under those conditions, the total activity of the recombinant protein was 30,650 U in the fermentation experiments, with an error of only 4.8%. The total activity of ASCHI61 increased 1.54-fold under the optimal conditions. Based on the results, ASCHI61 can be expressed more for hexaoligochitin production.
authors.affiliation: Graduate School of Biochemical Engineering and Department of Chemical Engineering, Ming Chi University of Technology, Taishan, New Taipei, Taiwan.
authors.forename: Chao-Lin
authors.initials: CL
authors.lastname: Liu
authors.forename: Tsung-Han
authors.initials: TH
authors.lastname: Lin
authors.forename: Ruey-Shin
authors.initials: RS
authors.lastname: Juang
mesh_terms: D061249:Batch Cell Culture Techniques / Q000379:methods / Q000592:standards; D002686:Chitin / Q000737:chemistry / Q000378:metabolism; D002688:Chitinases / Q000096:biosynthesis* / Q000378:metabolism; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism; D005285:Fermentation; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015203:Reproducibility of Results
chemical_list: D011994:Recombinant Proteins; D002686:Chitin; D002688:Chitinases
keywords: ASCHI61; Aeromonas schubertii chitinase of 61 kDa; CCA; CCD; Chitinase; GlcNAc; Hexaoligochitin; IPTG; N-acetyl-d-glucosamine; Optimization; RSM; Response surface methodology; central composite design; colloid chitin azure; isopropyl-β-d-thiogalactopyranoside; response surface methodology